Method for proliferation of antigen-specific T cells

Abstract

The present invention relates to an in vitro method for priming genetically modified T cells suitable for administration to a patient having a tumor. The invention is also directed to the composition obtained by the method and uses thereof.

The invention claimed is: 
     
       1. An in vitro method for priming of genetically modified antigen specific CD4+ and/or CD8+ T cells suitable for administration to a patient having a tumor, said method comprising co-culturing tumor antigen receptor expressing target T cells from the patient to be treated, mature dendritic cells, anti-CD3 antibodies and lymphocytes that have been sensitized against MHC class I and/or MHC class II antigens on antigen presenting cells (APCs), wherein the sensitized lymphocytes are obtained by culturing non-proliferating APCs from a first healthy donor with peripheral blood mononuclear cells (PMBCs) from a second healthy donor. 
     
     
       2. The method according to  claim 1 , wherein the tumor antigen receptors are selected from T-cell receptors (TCRs) and chimeric antigen receptors (CARs). 
     
     
       3. The method according to  claim 1 , wherein said non-proliferating antigen presenting cells are selected from the group consisting of PBMCs and monocyte derived dendritic cells. 
     
     
       4. The method according to  claim 1 , wherein the mature dendritic cells are obtained by first culturing monocytes in a composition comprising GM-CSF and IL-4 for about 1-7 days to obtain immature dendritic cells and subsequently adding a second composition that enables the immature dendritic cells to become mature dendritic cells by culturing for at least about 12 hours. 
     
     
       5. The method according to  claim 4 , wherein the second composition comprises TNF alfa, IL-1 beta, interferon gamma, interferon alpha or interferon beta and a TLR3 ligand. 
     
     
       6. The method according to  claim 4 , wherein the second composition comprises TNF alfa, interferon gamma, a TLR 3 ligand and/or a TLR 4 ligand, and a TLR7 agonist and/or a TLR 8 agonist. 
     
     
       7. The method according to  claim 6 , wherein the TLR 3 ligand is poly-1:C and the TLR 8 agonist is R848. 
     
     
       8. The method according to  claim 1 , wherein the cells are cultured for about 4 to 20 days. 
     
     
       9. The method according to  claim 1 , wherein exogenous IL-2, IL-7, IL-15, anti-IL-4 and/or IL-21 are added to the cell culture. 
     
     
       10. The method according to  claim 1 , wherein the primed antigen specific CD4+ and/or CD8+ T cells are restimulated by culturing said cells together with new dendritic cells, anti-CD3 antibodies, new sensitized allogeneic lymphocytes and optionally addition of exogenous IL-2, IL-7, IL-15, anti-IL-4 and/or IL-21 to the cell culture. 
     
     
       11. The method according to  claim 4 , wherein the TLR3 ligand comprises poly-1:C. 
     
     
       12. The method according to  claim 1 , wherein the non-proliferating APCs are autologous with respect to the mature dendritic cells. 
     
     
       13. The method according to  claim 1 , wherein the non-proliferating APCs are irradiated peripheral blood mononuclear cells (PMBCs).